title: Application of ultra-performance liquid chromatography/tandem mass spectrometry for the measurement of vitamin D in foods and nutritional supplements.
abstract: Ultra-performance liquid chromatography (UPLC)/MS/MS was applied to measure vitamin D in various foods and nutritional supplements. The run-time of the chromatographic separation was cut from 20 min in HPLC/MS/MS to 10 min in UPLC/MS/MS, while equal or better separation efficiency was achieved to deal with complex food matrixes. Under the optimized conditions, all the previtamins of vitamin D3, D2, and isotope-labeled vitamin D3 were baseline-separated from their corresponding vitamins. It was also demonstrated that many sterol isomers in complex food matrixes that interfere in the analysis could be well-separated from the analytes. Accuracy of this method was evaluated by analysis of NIST SRM 1849 infant formula reference material. With eight replicates, the average vitamin D3 concentration was 0.251 +/- 0.012 mg/kg, an excellent agreement with the certified value of 0.251 +/- 0.027 mg/kg. In addition, spike recovery from a commercial infant formula matrix was in the range of 100 to 108% for both vitamins D3 and D2 at three spike concentration levels. The spike recovery for an even more complex matrix, pet food, was 101-105%. LOQ values were 0.026 and 0.033 IU/g, or 0.086 and 0.11 IU/mL in solution, for vitamins D3 and D2, respectively. The dynamic range had three orders of magnitude, which made the method flexible and useful to deal with the wide concentration range of vitamin D in various samples. The method was robust based on the results of changing the parameters of LC separation and MS measurement. This accurate and reliable vitamin D method increased instrument efficiency and analysis productivity significantly.
authors.affiliation: Covance Laboratories Inc., 3301 Kinsman Blvd, Madison, WI 53704, USA. min.huang@covance.com
authors.forename: Min
authors.initials: M
authors.lastname: Huang
authors.forename: Doug
authors.initials: D
authors.lastname: Winters
mesh_terms: D002762:Cholecalciferol / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D019587:Dietary Supplements / Q000032:analysis*; D004872:Ergocalciferols / Q000032:analysis; D005504:Food Analysis / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D007223:Infant; D041943:Infant Formula / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000706:statistics & numerical data; D014807:Vitamin D / Q000032:analysis*
chemical_list: D004872:Ergocalciferols; D014807:Vitamin D; D002762:Cholecalciferol